B. SOW_OpAns_13Jun25 (6).pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

STATEMENT OF WORK (SOW)

(SERVICES)

GENERAL INFORMATION

Title of Project:

Bioanalytical analyses of steroid concentrations in blood samples.

Statement of Need and Purpose:

The purpose of this acquisition is to analyze steroid concentrations in blood samples using current LC-MS/MS methodology. OpAns’ steroid panel includes 14 compounds of interest in support of our study, characterizing the impacts of menopause stage, sex hormones, and sex chromosomes on brain aging and disease.

Background Information and Objective:

Biological sex influences risk for neurodegenerative diseases (NDDs), with Alzheimer’s disease occurring more often in females and Parkinson’s disease occurring more often in males. Here, we propose to investigate how hormonal fluctuations (associated with menopause and andropause) and sex chromosome dynamics shape both sex differences and within-sex variability in brain aging and NDD risk. We will generate new blood-based endocrine measures and integrate these data with existing intramural and published single-nucleus RNA sequencing (snRNA-seq) datasets from various brain regions to examine how menopause stage, sex hormone concentrations, and sex chromosome loss contribute brain aging in specific brain cell types. Specifically, we will measure: 1) blood steroid hormones using high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) performed by OpAns, LLC;

and 2) blood glycoproteins (FSH and AMH) using Enzyme-Linked Immunosorbent Assays (ELISAs) and an associated plate reader from Mesoscale Diagnostics, LLC. By sampling males and females across the reproductive aging period – including the menopause transition – this work can illuminate how hormonal fluctuations due to reproductive aging contribute to changes in brain gene expression in ways that may impact susceptibility to NDDs. Our findings will enhance our understanding of how sex-related mechanisms and aging interact to influence brain health, potentially revealing novel therapeutic targets for NDDs.

Period of Performance:

One year from date of post-award of services

SCOPE OF WORK

General Requirements:

• This proposal has been prepared with the assumption that all samples will be analyzed at the same time. If fewer than 50 samples are requested for analysis (either initial analyses or repeat analyses due to required dilutions), a minimum batch fee corresponding to the analysis of 50 samples applies. Extrapolated concentrations can be provided for no additional fee.

• The following will be provided once analysis is complete:

o An Excel data table minimally containing sample ID, matrix, and measured concentration o A formal report minimally containing standard and QC performance, sample concentrations, and a brief method summary (example chromatograms can be provided for $7200. This cost has been excluded from the cost schedule).

Specific Requirements:

OpAns’ steroid panel includes the following compounds over the following calibration ranges and utilizes an extraction volume of 200 μL for blood (up to 500 μL per sample would be ideal to account for re-analyses, if needed):

Aldosterone: 10 -10,000 pg/mL Androstenedione: 20 - 20,000 pg/mL Corticosterone: 200 - 200,000 pg/mL Cortisol: 600 - 600,000 pg/mL Cortisone: 200 - 200,000 pg/mL Dehydroepiandrosterone (DHEA): 600 - 600,000 pg/mL Deoxycorticosterone (DOC): 6 - 6,000 pg/mL 11-Deoxycortisol: 20 - 20,000 pg/mL Dihydrotestosterone (DHT): 60 - 60,000 pg/mL Estradiol: 10-10,000 pg/mL Estrone: 10-10,000 pg/mL 17α-OH-Progesterone: 60 - 60,000 pg/mL Progesterone: 20 - 20,000 pg/mL Testosterone: 10-10,000 pg/mL Extraction of lower volumes due to insufficient sample results in an upward shift of the above ranges that is inversely proportional to the sample volume extracted. Occasionally, low concentration standards need to be excluded due to a lack of instrument sensitivity. Blood concentrations will be provided in units of pg/mL. It should be noted that concentration ranges listed earlier correspond to plasma and sometimes need to be adjusted upward because of background chromatographic signal originating from the matrix (e.g. blood).

The liquid-liquid extraction methodology used to analyze samples for the above steroids has been applied to various biological matrices but has not been applied to all combinations of species and matrix. Occasionally, challenges such as chromatographic interferences or sample composition-related extraction issues are encountered. These challenges can hinder data interpretation for some mass channels or require the use of additional sample extraction cleanup using solid phase extraction. The cost for solid phase extraction plates, if needed, are invoiced at $400/plate (up to 96 wells). Reference standards (analytes and internal standards) and HPLC columns are included in the sample analysis cost.

This proposal has been prepared with the assumption that all samples will be analyzed at the same time.

If fewer than 50 samples are requested for analysis (either initial analyses or repeat analyses due to required dilutions), a minimum batch fee corresponding to the analysis of 50 samples applies. Extrapolated concentrations can be provided for no additional fee.

LEVEL OF EFFORT

The vendor will analyze and procure data as soon as practicable.

GOVERNMENT RESPONSIBILITIES

The government will provide vendor with an appropriate volume of blood, ~500ul would be optimal.

DELIVERY OR DELIVERABLES

Timely submission of analytical results are essential to successful completing this requirement.

REPORTING REQUIREMENTS

The following will be provided once analysis is complete:

o An Excel data table minimally containing sample ID, matrix, and measured concentration o A formal report minimally containing standard and QC performance, sample concentrations, and a brief method summary (example chromatograms can be provided for $7200. This cost has been excluded from the cost schedule).

OTHER CONSIDERATIONS

Travel:

Not applicable

Key Personnel:

Not applicable

Information System Security Plan:

Not Applicable

Data Rights:

Not Applicable

Section 508—Electronic and Information Technology Standards:

The contractor shall comply with Section 508 of the Rehabilitation Act (29 U.S.C. § 794d), as amended by the Workforce Investment Act of 1998 (P.L. 105-220), August 7, 1998.

Publications and Publicity:

Not Applicable

Confidentiality of Information:

Not Applicable